Frequency division multiplex HPLC-MS for simultaneous analyses.
Multiplex high-performance liquid chromatograph-mass spectrometry (HPLC-MS), in which multiple HPLCs and one MS are hyphenated, is an approach for high throughput analysis in HPLC-MS. A general multiplex HPLC-MS method employs a column-switching technology, and only one HPLC is connected to one MS at a time. In the present study, we propose a novel multiplex HPLC-MS system for simultaneous HPLC-MS analyses. In this study, multiple HPLCs are hyphenated with one MS without a column-switching mechanism, and a mixed-chromatogram is observed by the MS. Here, we employ a frequency division multiplexing (FDM) technique used in communication engineering to extract any chromatogram from the mixed-chromatogram. When a modulator (chopper or ion-gate type) is set between each ion source and the MS, each modulator blocks each sample stream with an individual frequency. In theory, each chromatogram can be extracted from the mixed-chromatogram via a signal processing based on a Fourier transform (FT), frequency-based signal extraction, and reversed FT. In the actual experiment, two HPLCs are hyphenated with one MS (2HPLC-1MS). The use of chopper type modulators leads to the extraction and restoration of each chromatogram from the mixed-chromatogram. However, each restored-chromatogram involves signal interference. On the other hand, the ion-gate modulation system successfully resulted in restored-chromatograms without interference. The potential of the novel multiplex HPLC-MS system based on FDM is confirmed with respect to the simultaneous and continuous analyses of plural samples.


Host and habitat filtering in seedling root-associated fungal communities: taxonomic and functional diversity are altered in 'novel' soils.
Climatic and land use changes have significant consequences for the distribution of tree species, both through natural dispersal processes and following management prescriptions. Responses to these changes will be expressed most strongly in seedlings near current species range boundaries. In northern temperate forest ecosystems, where changes are already being observed, ectomycorrhizal fungi contribute significantly to successful tree establishment. We hypothesised that communities of fungal symbionts might therefore play a role in facilitating, or limiting, host seedling range expansion. To test this hypothesis, ectomycorrhizal communities of interior Douglas-fir and interior lodgepole pine seedlings were analysed in a common greenhouse environment following growth in five soils collected along an ecosystem gradient. Currently, Douglas-fir's natural distribution encompasses three of the five soils, whereas lodgepole pine's extends much further north. Host filtering was evident amongst the 29 fungal species encountered: 7 were shared, 9 exclusive to Douglas-fir and 13 exclusive to lodgepole pine. Seedlings of both host species formed symbioses with each soil fungal community, thus Douglas-fir did so even where those soils came from outside its current distribution. However, these latter communities displayed significant taxonomic and functional differences to those found within the host distribution, indicative of habitat filtering. In contrast, lodgepole pine fungal communities displayed high functional similarity across the soil gradient. Taxonomic and/or functional shifts in Douglas-fir fungal communities may prove ecologically significant during the predicted northward migration of this species; especially in combination with changes in climate and management operations, such as seed transfer across geographical regions for forestry purposes.


Erythropoietic changes in rats after 2.45 GJz nonthermal irradiation.
The purpose of this study was to observe the erythropoietic changes in rats subchronically exposed to radiofrequency microwave (RF/MW) irradiation at nonthermal level. Adult male Wistar rats (N=40) were exposed to 2.45 GHz continuous RF/MW fields for 2 hours daily, 7 days a week, at 5-10 mW/cm2. Exposed animals were divided into four subgroups (n=10 animals in each subgroup) in order to be irradiated for 2, 8, 15 and 30 days. Animals were sacrified on the final irradiation day of each treated subgroup. Unexposed rats were used as control (N=24). Six animals were included into the each control subgroup. Bone marrow smears were examined to determine absolute counts of anuclear cells and erythropoietic precursor cells. The absolute erythrocyte count, haemoglobin and haematocrit values were observed in the peripheral blood by an automatic cell counter. The bone marrow cytogenetic analysis was accomplished by micronucleus (MN) tests. In the exposed animals erythrocyte count, haemoglobin and haematocrit were increased in peripheral blood on irradiation days 8 and 15. Concurrently, anuclear cells and erythropoietic precursor cells were significantly decreased (p < 0.05) in the bone marrow on day 15, but micronucleated cells' frequency was increased. In the applied experimental condition, RF/MW radiation might cause disturbance in red cell maturation and proliferation, and induce micronucleus formation in erythropoietic cells.


High Constitutive Cytokine Release by Primary Human Acute Myeloid Leukemia Cells Is Associated with a Specific Intercellular Communication Phenotype.
Acute myeloid leukemia (AML) is a heterogeneous disease, and this heterogeneity includes the capacity of constitutive release of extracellular soluble mediators by AML cells. We investigated whether this capacity is associated with molecular genetic abnormalities, and we compared the proteomic profiles of AML cells with high and low release. AML cells were derived from 71 consecutive patients that showed an expected frequency of cytogenetic and molecular genetic abnormalities. The constitutive extracellular release of 34 soluble mediators (CCL and CXCL chemokines, interleukins, proteases, and protease regulators) was investigated for an unselected subset of 62 patients, and they could be classified into high/intermediate/low release subsets based on their general capacity of constitutive secretion. FLT3-ITD was more frequent among patients with high constitutive mediator release, but our present study showed no additional associations between the capacity of constitutive release and 53 other molecular genetic abnormalities. We compared the proteomic profiles of two contrasting patient subsets showing either generally high or low constitutive release. A network analysis among cells with high release levels demonstrated high expression of intracellular proteins interacting with integrins, RAC1, and SYK signaling. In contrast, cells with low release showed high expression of several transcriptional regulators. We conclude that AML cell capacity of constitutive mediator release is characterized by different expression of potential intracellular therapeutic targets.


Negative frequency-dependent foraging behaviour in a generalist herbivore (Alces alces) and its stabilizing influence on food web dynamics.
Resource selection is widely appreciated to be context-dependent and shaped by both biological and abiotic factors. However, few studies have empirically assessed the extent to which selective foraging behaviour is dynamic and varies in response to environmental conditions for free-ranging animal populations. Here, we assessed the extent that forage selection fluctuated in response to different environmental conditions for a free-ranging herbivore, moose (Alces alces), in Isle Royale National Park, over a 10-year period. More precisely, we assessed how moose selection for coniferous versus deciduous forage in winter varied between geographic regions and in relation to (a) the relative frequency of forage types in the environment (e.g. frequency-dependent foraging behaviour), (b) moose abundance, (c) predation rate (by grey wolves) and (d) snow depth. These factors are potentially important for their influence on the energetics of foraging. We also built a series of food-chain models to assess the influence of dynamic foraging strategies on the stability of food webs. Our analysis indicates that moose exhibited negative frequency dependence, by selectively exploiting rare resources. Frequency-dependent foraging was further mediated by density-dependent processes, which are likely to be predation, moose abundance or some combination of both. In particular, frequency dependence was weaker in years when predation risk was high (i.e. when the ratio of moose to wolves was relatively low). Selection for conifers was also slightly weaker during deep snow years. The food-chain analysis indicates that the type of frequency-dependent foraging strategy exhibited by herbivores had important consequences for the stability of ecological communities. In particular, the dynamic foraging strategy that we observed in the empirical analysis (i.e. negative frequency dependence being mediated by density-dependent processes) was associated with more stable food web dynamics compared to fixed foraging strategies. The results of this study indicated that forage selection is a complex ecological process, varying in response to both biological (predation and moose density) and abiotic factors (snow depth) and over relatively small spatial scales (between regions). This study also provides a useful framework for assessing the influence of other aspects of foraging behaviour on the stability of food web dynamics.


Particle shape as revealed by spectral depolarization.
Through a series of numerical simulations we explore some scatter effects due to nonspherical particles. Specifically, we examine the link between the aspect ratio of randomly oriented, prolate spheroidal particles and the resulting linear depolarization of the scattered light in the forward and backscatter directions. The particular objective is to detect the presence of randomly oriented particles that have a systematic size and aspect ratio. Calculations show that the spectral behavior of the linear depolarization reveals the aspect ratio of the scattering particles. The concept is demonstrated using the size, shape, and refractive index of the spore form of Bacillus globigii (BG).


PET tumor metabolism in locally advanced breast cancer patients undergoing neoadjuvant chemotherapy: value of static versus kinetic measures of fluorodeoxyglucose uptake.
Changes in tumor metabolism from positron emission tomography (PET) in locally advanced breast cancer (LABC) patients treated with neoadjuvant chemotherapy (NC) are predictive of pathologic response. Serial dynamic [(18)F]-FDG (fluorodeoxyglucose) PET scans were used to compare kinetic parameters with the standardized uptake value (SUV) as predictors of pathologic response, disease-free survival (DFS), and overall survival (OS). Seventy-five LABC patients underwent FDG PET prior to and at midpoint of NC. FDG delivery (K(1)), FDG flux (K(i)), and SUV measures were calculated and compared by clinical and pathologic tumor characteristics using regression methods and area under the receiver operating characteristic curve (AUC). Associations between K(1), K(i), and SUV and DFS and OS were evaluated using the Cox proportional hazards model. Tumors that were hormone receptor negative, high grade, highly proliferative, or of ductal histology had higher FDG K(i) and SUV values; on an average, FDG K(1) did not differ systematically by tumor features. Predicting pathologic response in conjunction with estrogen receptor (ER) and axillary lymph node positivity, kinetic measures (AUC = 0.97) were more robust predictors than SUV (AUC = 0.84, P = 0.005). Changes in K(1) and K(i) predicted both DFS and OS, whereas changes in SUV predicted OS only. In multivariate modeling, only changes in K(1) remained an independent prognosticator of DFS and OS. Kinetic measures of FDG PET for LABC patients treated with NC accurately measured treatment response and predicted outcome compared with static SUV measures, suggesting that kinetic analysis may hold advantage of static uptake measures for response assessment.


Abundance and diversity of heterotrophic bacterial cells assimilating phosphate in the subtropical North Atlantic Ocean.
Microorganisms play key roles in the cycles of carbon and nutrients in the ocean, and identifying the extent to which specific taxa contribute to these cycles will establish their ecological function. We examined the use of (33)P-phosphate to identify heterotrophic bacteria actively involved in the cycling of phosphate, an essential inorganic nutrient. Seawater from the sub-tropical North Atlantic Ocean was incubated with (33)P-phosphate and analysed by microautoradiography to determine the proportion and diversity of the bacterial community-assimilating phosphate. Complementary incubations using (3)H-leucine and (3)H-thymidine were also conducted. We found that a higher proportion of total heterotrophic bacterial cells in surface water samples assimilated phosphate compared with leucine or thymidine. Bacteria from all of the phylogenetic groups we identified by CARD-FISH were able to assimilate phosphate, although the abundances of cells within each group did not scale directly with the number found to assimilate phosphate. Furthermore, a significantly higher proportion of Alphaproteobacteria, Gammaproteobacteria and Cytophaga-like cells assimilated phosphate compared with leucine or thymidine. Our results suggest that a greater proportion of bacterial cells in surface waters are actively participating in the biogeochemical cycling of phosphorus, and possibly other elements, than is currently estimated through the use of (3)H-leucine or (3)H-thymidine.


Nitrogen-Doped Nanoporous Carbon Membranes with Co/CoP Janus-Type Nanocrystals as Hydrogen Evolution Electrode in Both Acidic and Alkaline Environments.
Self-supported electrocatalysts being generated and employed directly as electrodes for energy conversion has been intensively pursued in the fields of materials chemistry and energy. Herein, we report a synthetic strategy to prepare freestanding hierarchically structured, nitrogen-doped nanoporous graphitic carbon membranes functionalized with Janus-type Co/CoP nanocrystals (termed as HNDCM-Co/CoP), which were successfully applied as a highly efficient, binder-free electrode in the hydrogen evolution reaction (HER). Benefited from multiple structural merits, such as a high degree of graphitization, three-dimensionally interconnected micro/meso/macropores, uniform nitrogen doping, well-dispersed Co/CoP nanocrystals, as well as the confinement effect of the thin carbon layer on the nanocrystals, HNDCM-Co/CoP exhibited superior electrocatalytic activity and long-term operation stability for HER under both acidic and alkaline conditions. As a proof-of-concept of practical usage, a 5.6 cm × 4 cm × 60 μm macroscopic piece of HNDCM-Co/CoP was prepared in our laboratory. Driven by a solar cell, electroreduction of water in alkaline conditions (pH 14) was performed, and H2 was produced at a rate of 16 mL/min, demonstrating its potential as real-life energy conversion systems.


Genotype at the -174G/C polymorphism of the interleukin-6 gene is associated with common carotid artery intimal-medial thickness: family study and meta-analysis.
Studies in unrelated individuals have produced conflicting findings concerning the putative association between the interleukin-6 (IL-6) -174G/C polymorphism and carotid intimal-medial thickness (IMT). We have used a family-based genetic association design to assess the heritability of carotid IMT and to investigate the hypothesized association of carotid IMT with the IL-6 to -174G/C polymorphism. We studied 854 members of 224 white British families. The heritability of carotid IMT was determined using Multipoint Engine for Rapid Likelihood Inference. Genetic association analyses were carried out using ANOVA and family-based tests of association implemented in Quantitative Transmission Disequilibrium Test. A meta-analysis of previous studies of the association was conducted to place our result in context. The heritability of carotid IMT was 24%. Under a recessive model (GG+GC versus CC), there was significant evidence of association between IL-6 to the -174G/C genotype and adjusted log(e) maximal carotid IMT (F=5.469; P=0.02). Family-based analyses using Quantitative Transmission Disequilibrium Test showed no evidence of population stratification as a cause of the observed association (chi2(1)=0.469; P=0.4934). The CC genotype was associated with a 4.8% increase in maximal carotid IMT and accounted for 0.6% of the observed variation in the trait, which is equivalent to 2.5% of the heritable component. A meta-analysis of the present and 2 previous large studies, which enrolled a total of 2930 subjects, confirmed the recessive effect of the C allele on carotid IMT (P=0.0014). The genotype at the IL-6 to -174G/C polymorphism is associated with common carotid artery IMT, although the size of the genetic effect is small.


Spinal navigation for posterior instrumentation of C1-2 instability using a mobile intraoperative CT scanner.
OBJECTIVE Spinal navigation techniques for surgical fixation of unstable C1-2 pathologies are challenged by complex osseous and neurovascular anatomy, instability of the pathology, and unreliable preoperative registration techniques. An intraoperative CT scanner with autoregistration of C-1 and C-2 promises sufficient accuracy of spinal navigation without the need for further registration procedures. The aim of this study was to analyze the accuracy and reliability of posterior C1-2 fixation using intraoperative mobile CT scanner-guided navigation. METHODS In the period from July 2014 to February 2016, 10 consecutive patients with instability of C1-2 underwent posterior fixation using C-2 pedicle screws and C-1 lateral mass screws, and 2 patients underwent posterior fixation from C-1 to C-3. Spinal navigation was performed using intraoperative mobile CT. Following navigated screw insertion in C-1 and C-2, intraoperative CT was repeated to check for the accuracy of screw placement. In this study, the accuracy of screw positioning was retrospectively analyzed and graded by an independent observer. RESULTS The authors retrospectively analyzed the records of 10 females and 2 males, with a mean age of 80.7 ± 4.95 years (range 42-90 years). Unstable pathologies, which were verified by fracture dislocation or by flexion/extension radiographs, included 8 Anderson Type II fractures, 1 unstable Anderson Type III fracture, 1 hangman fracture Levine Effendi Ia, 1 complex hangman-Anderson Type III fracture, and 1 destructive rheumatoid arthritis of C1-2. In 4 patients, critical anatomy was observed: high-riding vertebral artery (3 patients) and arthritis-induced partial osseous destruction of the C-1 lateral mass (1 patient). A total of 48 navigated screws were placed. Correct screw positioning was observed in 47 screws (97.9%). Minor pedicle breach was observed in 1 screw (2.1%). No screw displacement occurred (accuracy rate 97.9%). CONCLUSION Spinal navigation using intraoperative mobile CT scanning was reliable and safe for posterior fixation in unstable C1-2 pathologies with high accuracy in this patient series.


Additional contributions to taxonomy, nomenclature and biogeography of the Turkish Crataegus (Rosaceae) taxa.
Crataegusazarolus L. has a wide distribution pattern from the western Mediterranean coasts to the eastern parts of Iran with several varieties adapted to local climatic conditions. Crataegusazarolusvar.senobaaensisvar. nov. is described as a new variety from southeast Turkey with characteristic deep leaf sinuses, mostly 3-4 pairs of lobes and leaves ovate-oblong in outline. Two varieties of the species are accepted under the name of Crataegusazarolus and the correct names are published here. CrataegusmonogynaJacq.var.odemisiivar. nov. is described from İzmir, in the western part of Turkey. This new variety is distinguished by its orange fruit colour. An outstanding disjunct distribution pattern has been discovered for the recently described species, Crataegusyaltirikii Dönmez. Updated descriptions and infraspecific identification keys for Crataegusazarolus and Crataegusmonogyna are given and pictures and distribution data for the new taxa are also supplied.


Bubble over the head: Adeloye-Odeku disease in a Turkish child-case report.
Adeloye-Odeku disease is composed of dermoid cyst over anterior fontanelle, first described in 1971. We present an 11-year-old girl with a soft, fluctuant swelling over bregma. The lesion content was isointense to cerebrospinal fluid on both T2W and FLAIRW images. There was a lytic area under the lesion, seen on CT. The lesion was totally excised. Histopathology confirmed the diagnosis. The case was unique; because a fibrous band was observed extending to superior sagittal sinus and it has never been reported before. Although lesions are sub-aponeurotic, because of this kind of fibrous band, a thorough examination with neuroimaging tools is very important for planning of surgery.


Mesenchymal Stem Cells injection in hip osteoarthritis: preliminary results.
Osteoarthritis will become even more common in the near future since the average life span is steadily growing. Pain and loss of function are the main complaints reported by patients, inevitably leading towards a worsened daily life performance. New modern techniques have been developed with advanced cell based therapies. Mesenchymal stem cells (MSC) have the inner ability to mature into different types of cells depending on the stimuli they undergo. This technique has already been proven successful in the knee and, with this retrospective study, we would like to assess its feasibility in the hip joint. 6 consecutive patients affected by hip osteoarthritis were treated by intra-articular injection of autologous adipose-derived MSC between June 2017 and June 2018. Our study included only patients with constant hip pain resistant conservative treatment and OA graded 0-2 on the Tonnis grading scale. All 6 patients were evaluated in the preoperative setting and at the 6 months post-operative mark. The HHS showed an improvement from the pre-operative baseline mean value of 67.2±3.4 to the 84.6±6.3 post-operative value. Moreover, the WOMAC score dropped from a baseline score of 36.3±4.7 to 19.8±3.4 at 6 months' post-op follow up visit. MSC Lipogems is a fairly easy technique. No adverse effects were recorded in our experience. Preliminary results showed a positive outcome according to all the grading systems used in this study even though a longer follow up is needed to validate this technique.


Tasty THC: Promises and Challenges of Cannabis Edibles.
Food products containing cannabis extract (edibles) have emerged as a popular and lucrative facet of the legalized market for both recreational and medicinal cannabis. The many formulations of cannabis extracts used in edibles present a unique regulatory challenge for policy makers. Though edibles are often considered a safe, discreet, and effective means of attaining the therapeutic and/or intoxicating effects of cannabis without exposure to the potentially harmful risks of cannabis smoking, little research has evaluated how ingestion differs from other methods of cannabis administration in terms of therapeutic efficacy, subjective effects, and safety. The most prominent difference between ingestion and inhalation of cannabis extracts is the delayed onset of drug effect with ingestion. Consumers often do not understand this aspect of edible use and may consume a greater than intended amount of drug before the drug has taken effect, often resulting in profoundly adverse effects. Written for the educated layperson and for policy makers, this paper explores the current state of research regarding edibles, highlighting the promises and challenges that edibles present to both users and policy makers, and describes the approaches that four states in which recreational cannabis use is legal have taken regarding regulating edibles.


'High-tech' home care: overview of professional care in patients on home parenteral nutrition and implications for nursing care.
The aim of this study is to describe the quality, quantity and content of care given to home parenteral nutrition-dependent patients by various professionals in the Netherlands and to detect potential shortcomings. Home parenteral nutrition is a lifesaving treatment for patients who cannot eat or cannot eat sufficiently. Very little is known about follow-up care after hospital stay. Yet this is an important aspect of care as patients must cope with high-tech skills at home. Also, complications and psychosocial complaints can occur. Survey. A patient questionnaire was used to assess contacts with professionals and possible shortcomings in care. Nursing files from home parenteral nutrition nurses were reviewed for information from all contact moments. Home parenteral nutrition nurses and home care teams were interviewed to assess nursing care and to detect bottlenecks. The nutrition support team was primarily responsible for the home parenteral nutrition care. Physical complaints like abdominal pain or nausea and venous access problems like fever were discussed most often. Patients were satisfied about the nutrition support team, but both the patients and the home parenteral nutrition nurses reported that relatively little attention was paid to psychosocial problems. Furthermore, the included patients visited their General Practitioner 11 times per year on average (range 1-104). Patients experienced a bottleneck with respect to the general practitioner's knowledge of home parenteral nutrition-related matters. Home parenteral nutrition patients visit the nutrition support team and their general practitioner most frequently and much attention is paid to medical and physical problems. Psychosocial problems, however, were only discussed in a minority of patients, and this was experienced as a shortcoming. Relevance to clinical practice. Both the patients and the home parenteral nutrition nurses reported that relatively little attention was paid to psychosocial problems. Patients' concerns must be integrated in contact moments to enhance Quality of Life.


Life-Cycle Assessment of Gingko-Wood Three-Dimensional Membrane for Wastewater Treatment.
Forest is one of nature's most generous gifts to human beings, providing materials and shelters for all living beings with over 30% global land coverage. Apart from being sustainable, biodegradable, and renewable, wood is also extremely fascinating from the application aspect, with numerous advantages including hierarchical and macroporous structure, excellent mechanical performance, and versatile chemistry. The macroporous structure of wood is comprised of numerous long, partially aligned channels along the growth direction. This structure is suitable for a range of emerging applications, especially as a separation/membrane material. In this research, the potentiality of Gingko biloba (Gb) wood in the remediation of wastewater, contaminated with methylene blue (MB), a dye found in the industrial waters, was investigated. We report a macroporous, three-dimensional (3D) Gb-wood membrane decorated with palladium nanoparticles (Pd NPs) for efficient wastewater treatment. The efficiency of the Pd NPs/Gb-wood membrane to remove MB from a flowing aqueous solution was demonstrated. The wastewater treatment rate of the 3D Pd NPs/Gb-wood membrane can reach 0.5 L/min, with a high MB removal efficiency (>99.9%). The 3D Gb-wood macroporous membrane with partially aligned channels exhibits promising results for water treatment and is applicable for an even wider range of separation applications. In addition, the benefit of this 3D-wood membrane system for wastewater treatment was evaluated against the potential impacts on the environment and human health by employing the life-cycle assessment (LCA) approach. The LCA was carried out using the Gabi-education version with the gate-to-grave approach, including industrial wastewater, 3D-wood membrane, and electricity consumption using CML (Centrum VoorMilieukunde Leiden). From the LCA, it can be observed that wastewater filtration using this membrane exhibited a better environmental footprint due to the improved performance of the membrane in treating a higher volume of the permeate. Therefore, this filtration system had outweighed the additional environmental impact of the wastewater treatment process. The energy demand was identified as the main environmental hotspot in the LCA analysis. The analysis revealed that the energy source for electricity generation had a significant influence on the overall sustainability of this system. Additionally, the wood itself, a naturally abundant and eco-friendly material, presented zero environmental hazard to the environment during the filtration process. The experimental and environmental impact results indicate that Gb-wood can be employed as a natural and eco-friendly adsorbent material for the removal of waste from aqueous solutions.


Cell cycle-dependent phosphorylation of Rad53 kinase by Cdc5 and Cdc28 modulates checkpoint adaptation.
In budding yeast the evolutionarily conserved checkpoint response varies in its sensitivity to DNA damaging agents through the cell cycle. Specifically, higher amounts of damage are needed to activate the downstream checkpoint kinase Rad53 in S-phase cells. We examined here whether phosphorylation of Rad53 itself by cell cycle-dedicated kinases regulates Rad53 activation. We found that during unperturbed growth Rad53 exhibits a small phosphorylation-dependent electrophoretic mobility shift in G(2), M and G(1) phases of the cell cycle that is lost in S phase. We show that Rad53 is phosphorylated in vitro by Cdc5, a mitotic Polo-like kinase, and by the yeast cyclin-dependent kinase, Cdc28. Consistently, the cell cycle-dependent Rad53 mobility shift requires both Cdc5 and Cdc28 activities. We mapped the in vitro targeted phosphorylation sites by mass spectrometry and confirmed with mass spectroscopy that serines 774, 789 and 791 within Rad53 are phosphorylated in vivo in M-phase arrested cells. By creating nonphosphorylatable mutations in the endogenous RAD53 gene, we confirmed that the CDK and Polo kinase target sites are responsible for the observed cell cycle-dependent shift in protein mobility. The loss of phospho-acceptor sites does not interfere with Rad53 activation but accelerates checkpoint adaptation after induction of a single irreparable double-strand break. We thus demonstrate that cell cycle-dependent phosphorylation can fine-tune the response of Rad53 to DNA damage.


Babesia divergens-like organisms from free-ranging chamois (Rupicapra r. rupicapra) and roe deer (Capreolus c. capreolus) are distinct from B. divergens of cattle origin - an epidemiological and molecular genetic investigation.
In 2005 and 2006, three adult female chamois (Rupicapra r. rupicapra) were found dead with signs of acute babesial infection in the eastern Swiss Alps. PCR on DNA extracted from blood or spleen of the carcasses revealed sequence identity of the amplified part of the 18S rRNA gene with GenBank entries attributed to Babesia divergens of cattle origin or B. capreoli of wild ruminant origin which have never been described before in this region. Examination of 424 blood samples from 314 head of cattle from this area by IFAT, microscopy and PCR provided no evidence for babesial infection. Six of 887 ticks collected from cattle were PCR-positive, and sequencing revealed Babesia sp. genotype EU1 in five and B. divergens/B. capreoli in one of them. A Babesia isolate of chamois, two isolates of roe deer from the same region and one isolate of a roe deer from the north-western Swiss Alps were genetically compared with two Swiss B. divergens isolates of cattle origin by analysing the genomic rDNA locus. Whereas the near full length sequences of the 18S rRNA gene were virtually identical among all six isolates (>99.4% identity), distinct differences between the two isolates from cattle on the one hand and the four isolates from free-ranging ruminants on the other hand were observed in the sequences of the internal transcribed spacers 1 and 2 (ITS1, ITS2) and part of the 28S rRNA gene. These results indicate that, albeit genetically very closely related, these babesial organisms from cattle and from free-ranging ruminants indeed are distinguishable organisms with different host specificities, and they support the use of the discrete species name B. capreoli for the B. divergens-like organisms from chamois and roe deer.


Erythropoietin producing cells for potential cell therapy.
Anemia is an inevitable outcome of chronic renal failure due to the kidney's decreased ability to produce erythropoietin (EPO). We examined the feasibility of isolating and expanding EPO-producing cells for cell-based therapy. Renal cells from 7- to 10-day-old mice were culture-expanded. The cells at each subculture stage were characterized for EPO expression, using immunocytochemistry, FACS, and Western Blot analysis, with EPO-specific antibodies. To assess the levels of EPO expression, cells incubated under normoxic and hypoxic conditions were analyzed. Immunocytochemical analysis of the cultured renal cells expressed EPO at each subculture stage (P1-P3). Western Blot analysis of the detergent-solubilized cell extracts detected EPO (34 kDa) protein in the kidney cells of all passages tested. These results demonstrate that EPO-producing renal cells can be grown and expanded in culture. The cells stably expressed EPO at multiple subculture stages and they are able to form tissue in vivo. This study shows that EPO-producing cells may be used as a potential treatment option for anemia caused by chronic renal failure.


Incremental prognostic value of cardiac computed tomography in coronary artery disease using CONFIRM: COroNary computed tomography angiography evaluation for clinical outcomes: an InteRnational Multicenter registry.
Large multicenter studies validating the prognostic value of coronary computed tomographic angiography (CCTA) and left ventricular ejection fraction (LVEF) are lacking. We sought to confirm the independent and incremental prognostic value of coronary artery disease (CAD) severity measured using 64-slice CCTA over LVEF and clinical variables. A large international multicenter registry (CONFIRM Registry) was queried, and CCTA patients with LVEF data on CCTA were screened. Patients with a history of myocardial infarction, coronary revascularization, or cardiac transplantation were excluded. The National Cholesterol Education Program-Adult Treatment Panel III risk was calculated for each patient, and CCTA was evaluated for CAD severity (normal, nonobstructive, non-high-risk, or high-risk CAD) and LVEF <50%. Patients were followed for an end point of all-cause mortality; 27 125 patients underwent CCTA at 12 participating centers, with a total of 14 064 patients meeting the analysis criteria. Follow-up was available for 13 966 (99.3%) patients (mean follow-up of 22.5 months; 95% confidence interval, 22.3 to 22.7 months). All-cause mortality (271 deaths) occurred in 0.65% of patients without coronary atherosclerosis, 1.99% of patients with nonobstructive CAD, 2.90% of patients with non-high-risk CAD, and 4.95% for patients with high-risk CAD. Multivariable analysis confirmed that LVEF <50% (hazard ratio, 2.74; 95% confidence interval, 2.12 to 3.51) and CAD severity (hazard ratio,1.58; 95% confidence interval, 1.42 to 1.76) were predictors of all-cause mortality, and CAD severity had incremental value over LVEF and clinical variables. Our results demonstrate that CCTA measures of CAD severity and LVEF have independent prognostic value. Incorporation of CAD severity provides incremental value for predicting all-cause death over routine clinical predictors and LVEF in patients with suspected obstructive CAD.


The carboxyl terminal sequence of nucleolar protein B23.1 is important in its DNA polymerase alpha-stimulatory activity.
The protein B23 is a major nucleolar phosphoprotein comprising two isoforms, B23.1 and B23.2, which differ only in their carboxyl-terminal short sequences, the N-terminal 255 residues being identical in both forms. Both B23.1 and B23.2 stimulated immunoaffinity-purified calf thymus DNA polymerase alpha in a dose-dependent manner. The stimulatory effect of protein B23.1, the longer isoform, was found to be 2-fold greater than that of B23.2. Purified DNA polymerase alpha bound tightly to a protein B23.1-immobilized column, while it bound weakly to a protein B23.2-immobilized column. Surface plasmon resonance studies by BIAcore further showed that protein B23.1 bound to the DNA polymerase alpha-(dA).(dT) complex more tightly than did protein B23.2. The protein B23 isoforms appear to interact directly with the DNA polymerase alpha protein and not through the bound nucleic acid. These observations indicated that protein B23 physically bound to the DNA polymerase alpha and stimulated the enzyme activity. Product analyses showed that protein B23 greatly enhanced the reaction both in amount and length of product DNA, whereas it did not significantly alter the processivity of polymerization. In contrast, protein B23 effectively protected DNA polymerase alpha from heat inactivation. These results suggest that protein B23 stabilizes DNA polymerase alpha that is detached from product DNA, allowing the enzyme to be recruited for further elongation. Moreover, experiments using various C-terminal deletion mutants of protein B23 indicated that 12 amino acids at the C-terminal end of B23.1, which are absent in B23.2, may be essential for the full stimulation of the DNA polymerase alpha.


Neuroscience of human social interactions and adult attachment style.
Since its first description four decades ago, attachment theory (AT) has become one of the principal developmental psychological frameworks for describing the role of individual differences in the establishment and maintenance of social bonds between people. Yet, still little is known about the neurobiological underpinnings of attachment orientations and their well-established impact on a range of social and affective behaviors. In the present review, we summarize data from recent studies using cognitive and imaging approaches to characterize attachment styles and their effect on emotion and social cognition. We propose a functional neuroanatomical framework to integrate the key brain mechanisms involved in the perception and regulation of social emotional information, and their modulation by individual differences in terms of secure versus insecure (more specifically avoidant, anxious, or resolved versus unresolved) attachment traits. This framework describes how each individual's attachment style (built through interactions between personal relationship history and predispositions) may influence the encoding of approach versus aversion tendencies (safety versus threat) in social encounters, implicating the activation of a network of subcortical (amygdala, hippocampus, striatum) and cortical (insula, cingulate) limbic areas. These basic and automatic affective evaluation mechanisms are in turn modulated by more elaborate and voluntary cognitive control processes, subserving mental state attribution and emotion regulation capacities, implicating a distinct network in medial prefrontal cortex (mPFC), superior temporal sulcus (STS), and temporo-parietal junction (TPJ), among others. Recent neuroimaging data suggest that affective evaluation is decreased in avoidantly but increased in anxiously attached individuals. In turn, although data on cognitive control is still scarce, it points toward a possible enhancement of mental state representations associated with attachment insecurity and particularly anxiety. Emotion regulation strategies such as reappraisal or suppression of social emotions are also differentially modulated by attachment style. This research does not only help better understand the neural underpinnings of human social behavior, but also provides important insights on psychopathological conditions where attachment dysregulation is likely to play an important (causal) role.


Factors associated with selling price of cattle at livestock marts.
The objective of this study was to determine the factors associated with selling price of animals at livestock marts around Ireland. Data consisted of four distinct maturity categories: calves (2 to 84 days of age, n = 53 838); weanlings (6 to 12 months of age, n = 19 972); post-weanlings (12 to 36 months of age, n = 93 081) and cows (>30 months to 12 years of age, n = 94 839); sold through livestock marts between 2000 and 2008. Factors associated with animal price were determined within each maturity category separately using mixed models; random effects were mart, date of sale nested within mart, and herd of origin nested within year of sale. Mean selling price was €157, €580, €655 and €592 for calves, weanlings, post-weanlings and cows, respectively. The greatest prices were paid for singleton crossbred male calves, weanlings and post-weanlings from older dams. With the exception of the Aberdeen Angus, beef breeds and their crosses consistently received higher prices than their dairy counterparts across all four maturity categories; increased proportion of Belgian Blue and Charolais was associated with greater prices compared with other beef breeds. When live-weight was included in the multiple regression models the association between price and all factors regressed toward zero but most factors remained associated with price. The highest price was recorded in the spring months for calves, post-weanlings and cows, and in the autumn months for weanlings. Results from this study may be used to help farmers make more informed management decisions, as well as provide information for bio-economic models for evaluating alternative production systems or estimating economic values.


Requirement for a GTPase-activating protein in vesicle budding from the endoplasmic reticulum.
The binding and hydrolysis of guanosine triphosphate (GTP) by the small GTP-binding protein Sar1p is required to form transport vesicles from the endoplasmic reticulum (ER) in Saccharomyces cerevisiae. Experiments revealed that an interaction between Sar1p and the Sec23p subunit of an oligomeric protein is also required for vesicle budding. The isolated Sec23p subunit and the oligomeric complex stimulated guanosine triphosphatase (GTPase) activity of Sar1p 10- to 15-fold but did not activate two other small GTP-binding proteins involved in vesicle traffic (Ypt1p and ARF). Activation of GTPase was inhibited by an antibody to Sec23p but not by an antibody that inhibits the budding activity of the other subunit of the Sec23p complex. Also, activation was thermolabile in pure samples of Sec23p that were isolated from two independent sec23 mutant strains. It appears that Sec23p represents a new class of GTPase-activating protein because its sequence shows no similarity to any known member of this family.


Gaucher disease and bone manifestations.
Gaucher disease is a relatively rare metabolic disease caused by the inherited deficiency of the lysosomal enzyme glucocerebrosidase. Gaucher disease affects multiple organs, among which is the skeleton. Bone involvement occurs frequently in Gaucher disease, and is one of its most debilitating features, reducing the quality of life of patients. Bone status is an important consideration for treatment to ameliorate symptoms and reduce the risk of irreversible complications. We have conducted a systematic review of all the various aspects of Gaucher disease, focusing on different skeletal manifestations, pathophysiology of bone alterations, clinical symptoms, and current diagnostic and therapeutic approaches.


Structural and bonding patterns in gold clusters.
The study of gold cluster compounds originated from Malatesta's syntheses of tertiaryphosphine derivatives in the 1960s and was greatly extended between 1970 and 2000. Single crystal X-ray studies defined the major structural classes and led to the development of a theoretical model which accounted for their closed shell requirements in terms of their topological features and proved to be sufficiently flexible to be extended to related heteronuclear cluster compounds. Since the turn of the century the range of gold cluster compounds has been greatly extended by the study of organothiolato-gold cluster compounds. The structures of these compounds have revealed that the gold atoms combine with the organothiolato-ligands to generate a novel class of metallo-organothiolato-ligands which protect and stabilise the inner core of gold atoms. These developments originally suggested that the phosphine and organothiolato-clusters defined quite distinct classes of gold clusters, but recent structural and theoretical developments have reconciled many of these differences. This review summarises the structures of all the clusters of gold and suggests a theoretical model which effectively unites the broad structural properties of the two classes of compound. This model is based on the united atom model for diatomics developed by Mulliken and the compression co-ordinate is related to the interpenetration of icosahedral and cuboctahedral pseudo-spherical clusters. The predicted closed shell requirements agree well with the results of structural determinations.


New-generation oral anticoagulants for the prevention of stroke: implications for neurosurgery.
A new generation of oral anticoagulants, namely direct thrombin inhibitors and factor Xa inhibitors, have recently been approved for clinical use in patients with atrial fibrillation. These novel families of drugs have been shown to have favorable efficacy and safety profiles in multiple clinical settings, particularly in the prevention of atrial fibrillation-related stroke, and are likely to become part of everyday practice, making a crossover to neurosurgical patients inevitable. Concern has risen regarding the complexity of managing intracranial and intraspinal hemorrhages related to these drugs. This review aims to provide an update on the most recent advances in oral anticoagulant drug therapy from a neurosurgeon's perspective. We discuss current evidence for the use of these novel agents, their limitations, existing methods of drug-level monitoring, and controversies related to anticoagulation reversal. We also discuss specific topics such as anticoagulation resumption after intracranial or intraspinal bleeding, perioperative anticoagulant administration, and the possibility of combination with tissue plasminogen activator in the setting of acute ischemic stroke. A special focus is given to the incidence of intracranial and intraspinal hemorrhage associated with each drug.


FDA-approved selective estrogen receptor modulators inhibit Ebola virus infection.
Ebola viruses remain a substantial threat to both civilian and military populations as bioweapons, during sporadic outbreaks, and from the possibility of accidental importation from endemic regions by infected individuals. Currently, no approved therapeutics exist to treat or prevent infection by Ebola viruses. Therefore, we performed an in vitro screen of Food and Drug Administration (FDA)- and ex-US-approved drugs and selected molecular probes to identify drugs with antiviral activity against the type species Zaire ebolavirus (EBOV). From this screen, we identified a set of selective estrogen receptor modulators (SERMs), including clomiphene and toremifene, which act as potent inhibitors of EBOV infection. Anti-EBOV activity was confirmed for both of these SERMs in an in vivo mouse infection model. This anti-EBOV activity occurred even in the absence of detectable estrogen receptor expression, and both SERMs inhibited virus entry after internalization, suggesting that clomiphene and toremifene are not working through classical pathways associated with the estrogen receptor. Instead, the response appeared to be an off-target effect where the compounds interfere with a step late in viral entry and likely affect the triggering of fusion. These data support the screening of readily available approved drugs to identify therapeutics for the Ebola viruses and other infectious diseases. The SERM compounds described in this report are an immediately actionable class of approved drugs that can be repurposed for treatment of filovirus infections.


Neonatal corticosterone mitigates autoimmune neuropsychiatric disorders associated with streptococcus in mice.
Increased glucocorticoid concentrations have been shown to favor resilience towards autoimmune phenomena. Here, we addressed whether experimentally induced elevations in circulating glucocorticoids mitigate the abnormalities exhibited by an experimental model of Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcus (PANDAS). This is a pathogenic hypothesis linking repeated exposures to Group-A-beta-hemolytic streptococcus (GAS), autoantibodies targeting selected brain nuclei and neurobehavioral abnormalities. To persistently elevate glucocorticoid concentrations, we supplemented lactating SJL/J mice with corticosterone (CORT; 80 mg/L) in the drinking water. Starting in adolescence (postnatal day 28), developing offspring were exposed to four injections - at bi-weekly intervals - of a GAS homogenate and tested for behavioral, immunological, neurochemical and molecular alterations. GAS mice showed increased perseverative behavior, impaired sensorimotor gating, reduced reactivity to a serotonergic agonist and inflammatory infiltrates in the anterior diencephalon. Neonatal CORT persistently increased circulating glucocorticoids concentrations and counteracted these alterations. Additionally, neonatal CORT increased peripheral and CNS concentrations of the anti-inflammatory cytokine IL-9. Further, upstream regulator analysis of differentially expressed genes in the striatum showed that the regulatory effect of estradiol is inhibited in GAS-treated mice and activated in GAS-treated mice exposed to CORT. These data support the hypothesis that elevations in glucocorticoids may promote central immunomodulatory processes.


Residential proximity to major roadways and incident hypertension in post-menopausal women.
Living near major roadways has been associated with increased risk of cardiovascular morbidity and mortality, presumably from exposure to elevated levels of traffic-related air and/or noise pollution. This association may potentially be mediated through increased risk of incident hypertension, but results from prior studies are equivocal. Using Cox proportional hazards models we examined residential proximity to major roadways and incident hypertension among 38,360 participants of the Women's Health Initiative (WHI) Clinical Trial cohorts free of hypertension at enrollment and followed for a median of 7.9 years. Adjusting for participant demographics and lifestyle, trial participation, and markers of individual and neighborhood socioeconomic status, the hazard ratios for incident hypertension were 1.13 (95% CI: 1.00, 1.28), 1.03 (0.95, 1.11), 1.05 (0.99, 1.11), and 1.05 (1.00, 1.10) for participants living ≤50, >50-200, >200-400, and >400-1000 m vs >1000 m from the nearest major roadway, respectively (ptrend=0.013). This association varied substantially by WHI study region with hazard ratios for women living ≤50 m from a major roadway of 1.61 (1.18, 2.20) in the West, 1.51 (1.22, 1.87) in the Northeast, 0.89 (0.70, 1.14) in the South, and 0.94 (0.75, 1.19) in the Midwest. In this large, national cohort of post-menopausal women, residential proximity to major roadways was associated with incident hypertension in selected regions of the U.S. If causal, these results suggest residential proximity to major roadways, as a marker for air, noise and other traffic-related pollution, may be a risk factor for hypertension.


A second dose of a CFTR cDNA-liposome complex is as effective as the first dose in restoring cAMP-dependent chloride secretion to null CF mice trachea.
Phase I clinical trials have provided encouraging data suggesting that gene transfer could provide a treatment for cystic fibrosis (CF). However, for all the current viral and nonviral vectors used to deliver the cystic fibrosis transmembrane conductance regulator (CFTR) gene, the duration of CFTR expression is limited, necessitating a repeat dosing regimen to provide a long-term treatment. This study was performed to determine whether a second delivery of a CFTR cDNA-liposome complex could result in a similar level of functional CFTR expression observed after a single delivery and to assess whether the deliveries produced adverse inflammatory responses. CFTR functional expression was assessed by short circuit current measurements of tracheas taken from CF null mice (Cftrtm1Cam) treated with a CFTR cDNA-liposome complex in the upper airways. Mice receiving two deliveries of this complex, the second after the response to the first had declined, showed cAMP-stimulated chloride currents which were not significantly different from normal tracheas or tissues assayed after a single dose of the complex. This double treatment was well tolerated with no discernible inflammation of lung tissue.


Craniotomy flap osteomyelitis: a diagnostic approach.
Nine cases of suspected craniotomy flap osteomyelitis evaluated by combined bone and gallium scanning are presented. In six cases, the clinical data were inconclusive and evaluation by radionuclide imaging provided an accurate negative diagnosis. The other three cases considered positive by this technique were proven infected at subsequent exploration and flap removal. The use of radionuclide bone and gallium imaging should be considered in cases of possible craniotomy flap osteomyelitis.


Selective inhibition of plant serine hydrolases by agrochemicals revealed by competitive ABPP.
Organophosphate and -phosphonates and their thio derivatives are often used in agroindustry as herbicides and insecticides, but their potential off-targets in the plant are poorly investigated. Here, we use competitive activity-based protein profiling (ABPP) of serine hydrolases (SHs) to detect targets of these agrochemicals and other compounds in Arabidopsis thaliana. Using broad-range and specific probes, and by overexpression of various SHs in planta, we are able to confirm eight SH-compound interactions, including selective inhibition of carboxylesterase CXE12, prolyloligopeptidase, methylesterase MES2 and tripeptidyl peptidase TPP2. These observations can be used for the design of novel probes and selective inhibitors and may help to assess physiological effects of agrochemicals on crop plants.


Antibiotic treatment of children with unsuspected meningococcal disease.
Data from an earlier study suggest that patients with unsuspected meningococcal disease (UMD) cannot be differentiated easily from febrile children with viral syndromes on the basis of physical examinations or peripheral blood counts. Some children with meningococcal disease therefore are treated inadvertently as outpatients. To determine whether antibiotic therapy administered at the outpatient visit prevents complications, permanent sequelae, or death in children with UMD. We reviewed the medical records of patients younger than 20 years with invasive meningococcal disease at 7 pediatric referral centers from January 1, 1981, through December 31, 1996. Patients were considered to have UMD if they underwent evaluation and discharge as outpatients and if blood and/or cerebrospinal fluid cultures obtained at evaluation yielded Neisseria meningitidis. We compared the frequency of development of complications (meningitis, sepsis, and pericarditis), permanent sequelae (limb amputation, skin grafting, and persistent neurologic disability) or death between patients who did and did not receive antibiotics at the outpatient visits. Of 58 children with UMD, 19 (33%) received antibiotics and 39 (67%) did not. Complications occurred significantly less frequently in the antibiotic-treated group (7/19 [37%] vs 27/39 [69%]; odds ratio [OR], 0.26; 95% confidence interval [CI], 0.08-0.81; P = .03). There was no significant difference in death or permanent sequelae between groups (0/19 vs 3/39 [8%]; OR, 0; 95% CI, 0-2.61; P=.54). There was insufficient power, however, to exclude the possibility of a clinically meaningful difference between the groups with regard to these latter outcomes. Antibiotic administration to young patients with UMD at the time of the outpatient visit is associated with a reduction in complications from this disease. Although the routine use of antibiotics in febrile outpatients younger than 20 years cannot be advocated, empirical treatment should be considered in the setting of higher probability of meningococcal disease.


Evaluation of butorphanol, medetomidine and midazolam as a reversible narcotic combination in free-ranging African lions (Panthera leo).
To evaluate the effects of the combination butorphanol, medetomidine and midazolam (BMM) and its reversibility in lions. Prospective clinical trial. Thirty free-ranging lions, 10 male and 20 female, weighing 81-210 kg. Lions were immobilised with butorphanol mean 0.31 ± SD 0.034 mg kg(-1), medetomidine 0.052 ± 0.006 mg kg(-1), midazolam 0.21 ± 0.024 mg kg(-1) and hyaluronidase 1250 IU administered intramuscularly with a dart gun. Upon recumbency, physiological parameters and anaesthetic depth were monitored 10-15 minutes after darting (T1) and repeated every 10 minutes for a further 30 minutes (T2, T3, T4). Arterial blood gas analyses were performed at T1 and T4. At the end of the procedure, 45-60 minutes after initial darting, immobilisation was reversed with naltrexone 0.68 ± 0.082 mg kg(-1), atipamezole 0.26 ± 0.031 mg kg(-1), and flumazenil 0.0032 ± 0.0007 mg kg(-1) administered intravenously and subcutaneously. The BMM combination rapidly induced immobilisation and lateral recumbency was reached within 7.25 ± 2.3 minutes. Median induction score [scored 1 (excellent) to 4 (poor)] was 1.4 (range 1-2). Cardio-respiratory parameters were stable. Heart rate varied from 32 to 72 beats per minute, respiratory rate from 14 to 32 breaths minute(-1) and rectal temperature from 36.6 to 40.3 °C. No sudden arousals were observed. Arterial blood gas analyses revealed a mean pH of 7.33, PaCO(2) of 33 mmHg and PaO(2) of 87 mmHg. Mild to moderate hypoxemia was seen in four lions. Recovery was smooth and lions were walking within 4.4 ± 4.25 minutes. Median recovery score [scored 1 (excellent) to 4 (poor)] was 1.3 (range 1-2). The drug combination proved to be effective in immobilising free-ranging healthy lions of both sexes with minimal cardio-respiratory changes.


Learning curves, taking instructions, and patient safety: using a theoretical domains framework in an interview study to investigate prescribing errors among trainee doctors.
Prescribing errors are a major source of morbidity and mortality and represent a significant patient safety concern. Evidence suggests that trainee doctors are responsible for most prescribing errors. Understanding the factors that influence prescribing behavior may lead to effective interventions to reduce errors. Existing investigations of prescribing errors have been based on Human Error Theory but not on other relevant behavioral theories. The aim of this study was to apply a broad theory-based approach using the Theoretical Domains Framework (TDF) to investigate prescribing in the hospital context among a sample of trainee doctors. Semistructured interviews, based on 12 theoretical domains, were conducted with 22 trainee doctors to explore views, opinions, and experiences of prescribing and prescribing errors. Content analysis was conducted, followed by applying relevance criteria and a novel stage of critical appraisal, to identify which theoretical domains could be targeted in interventions to improve prescribing. Seven theoretical domains met the criteria of relevance: "social professional role and identity," "environmental context and resources," "social influences," "knowledge," "skills," "memory, attention, and decision making," and "behavioral regulation." From critical appraisal of the interview data, "beliefs about consequences" and "beliefs about capabilities" were also identified as potentially important domains. Interrelationships between domains were evident. Additionally, the data supported theoretical elaboration of the domain behavioral regulation. In this investigation of hospital-based prescribing, participants' attributions about causes of errors were used to identify domains that could be targeted in interventions to improve prescribing. In a departure from previous TDF practice, critical appraisal was used to identify additional domains that should also be targeted, despite participants' perceptions that they were not relevant to prescribing errors. These were beliefs about consequences and beliefs about capabilities. Specifically, in the light of the documented high error rate, beliefs that prescribing errors were not likely to have consequences for patients and that trainee doctors are capable of prescribing without error should also be targeted in an intervention. This study is the first to suggest critical appraisal for domain identification and to use interview data to propose theoretical elaborations and interrelationships between domains.


Harnessing the Power of SIRT1 and Non-coding RNAs in Vascular Disease.
Noncommunicable diseases (NCDs) contribute to a significant amount of disability and death in the world. Of these disorders, vascular disease is ranked high, falls within the five leading causes of death, and impacts multiple other disease entities such as those of the cardiac system, nervous system, and metabolic disease. Targeting the silent mating type information regulation 2 homolog 1 (Saccharomyces cerevisiae) (SIRT1) pathway and the modulation of micro ribonucleic acids (miRNAs) may hold great promise for the development of novel strategies for the treatment of vascular disease since each of these pathways are highly relevant to cardiac and nervous system disorders as well as to metabolic dysfunction. SIRT1 is vital in determining the course of stem cell development and the survival, metabolism, and life span of differentiated cells that are overseen by both autophagy and apoptosis. SIRT1 interfaces with a number of pathways that involve forkhead transcription factors, mechanistic of rapamycin (mTOR), AMP activated protein kinase (AMPK) and Wnt1 inducible signaling pathway protein 1 (WISP1) such that the level of activity of SIRT1 can become a critical determinant for biological and clinical outcomes. The essential fine control of SIRT1 is directly tied to the world of non-coding RNAs that ultimately oversee SIRT1 activity to either extend or end cellular survival. Future studies that can further elucidate the crosstalk between SIRT1 and non-coding RNAs should serve well our ability to harness the power of SIRT1 and non-coding RNAs for the treatment of vascular disorders.


Replication study of 10 genetic polymorphisms associated with coronary heart disease in a specific high-risk population with familial hypercholesterolemia.
Recent large association studies have revealed associations between genetic polymorphisms and myocardial infarction and coronary heart disease (CHD). We performed a replication study of 10 polymorphisms and CHD in a population with familial hypercholesterolemia (FH), individuals at extreme risk of CHD. We genotyped 10 polymorphisms in 2145 FH patients and studied the association between these polymorphisms and CHD in Cox proportional hazards models. We confirmed the associations between four polymorphisms and CHD, the rs1151640 polymorphism in the olfactory receptor family 13 subfamily G member 1 (OR13G1) gene (HR 1.14, 95% CI 1.01-1.28, P = 0.03), the rs11881940 polymorphism in the heterogeneous nuclear ribonucleoprotein U-like 1 (HNRPUL1) gene (HR 1.27, 95% CI 1.07-1.51, P = 0.007), the rs3746731 polymorphism in the complement component 1 q subcomponent receptor 1 (CD93) gene (HR 1.26, 95% CI 1.06-1.49, P = 0.01), and the rs10757274 polymorphism near the cyclin-dependent kinase N2A and N2B (CDKN2A and CDKN2B) genes (HR 1.39, 95% CI 1.15-1.69, P < 0.001). We confirmed previously found associations between four polymorphisms and CHD, but refuted associations for six other polymorphisms in our large FH population. These findings stress the importance of replication before genetic information can be implemented in the prediction of CHD.


Atmospheric correction of HJ-1 CCD imagery over turbid lake waters.
We have presented an atmospheric correction algorithm for HJ-1 CCD imagery over Lakes Taihu and Chaohu with highly turbid waters. The Rayleigh scattering radiance (Lr) is calculated using the hyperspectral Lr with a wavelength interval 1nm. The hyperspectral Lr is interpolated from Lr in the central wavelengths of MODIS bands, which are converted from the band response-averaged Lr calculated using the Rayleigh look up tables (LUTs) in SeaDAS6.1. The scattering radiance due to aerosol (La) is interpolated from La at MODIS band 869nm, which is derived from MODIS imagery using a shortwave infrared atmospheric correction scheme. The accuracy of the atmospheric correction algorithm is firstly evaluated by comparing the CCD measured remote sensing reflectance (Rrs) with MODIS measurements, which are validated by the in situ data. The CCD measured Rrs is further validated by the in situ data for a total of 30 observation stations within ± 1h time window of satellite overpass and field measurements. The validation shows the mean relative errors about 0.341, 0.259, 0.293 and 0.803 at blue, green, red and near infrared bands.


Interventions to Increase Physical Activity in Patients with COPD: A Comprehensive Review.
It is unknown how interventions aimed at increasing physical activity (PA), other than traditional pulmonary rehabilitation, are structured and whether they are effective in increasing PA in chronic obstructive pulmonary disease (COPD). The primary aim of this review was to outline the typical components of PA interventions in patients with COPD. This review followed the PRISMA guidelines. A structured literature search of relevant electronic databases from inception to April 2014 was undertaken to outline typical components and examine outcome variables of PA interventions in patients with COPD. Over 12000 articles were screened and 20 relevant studies involving 31 PA interventions were included. Data extracted included patient demographics, components of the PA intervention, PA outcome measures and effects of the intervention. Quality was assessed using the PEDro and CASP scales. There were 13 randomised controlled trials and three randomised trials (PEDro score 5-7/10) and four cohort studies (CASP score 5/10). Interventions varied in duration, number of participant/researcher contacts and mode of delivery. The most common behaviour change techniques included information on when and where (n = 26/31) and how (n = 22/31) to perform PA behaviour and self-monitoring (n = 18/31). Significant between-group differences post-intervention in favour of the PA intervention, compared to a control group or to other PA interventions, in one or more PA assessments were found in 7/16 studies. All seven studies used walking as the main type of PA/exercise. In conclusion, although the components of PA interventions were variable, there is some evidence that PA interventions have the potential to increase PA in patients with COPD.


Allelic variants of the Pseudomonas syringae type III effector HopZ1 are differentially recognized by plant resistance systems.
The bacterial plant pathogen Pseudomonas syringae depends on the type III secretion system and type III-secreted effectors to cause disease in plants. HopZ is a diverse family of type III effectors widely distributed in P. syringae isolates. Among the HopZ homologs, HopZ1 is ancient to P. syringae and has been shown to be under strong positive selection driven by plant resistance-imposed selective pressure. Here, we characterized the virulence and avirulence functions of the three HopZ1 alleles in soybean and Nicotiana benthamiana. In soybean, HopZ1 alleles have distinct functions: HopZ1a triggers defense response, HopZ1b promotes bacterial growth, and HopZ1c has no observable effect. In N. benthamiana, HopZ1a and HopZ1b both induce plant defense responses. However, they appear to trigger different resistance pathways, evidenced by two major differences between HopZ1a- and HopZ1b-triggered hypersensitive response (HR): i) the putative N-acylation sites had no effect on HopZ1a-triggered cell death, whereas it greatly enhanced HopZ1b-triggered cell death; and ii) the HopZ1b-triggered HR, but not the HopZ1a-triggered HR, was suppressed by another HopZ homolog, HopZ3. We previously demonstrated that HopZ1a most resembled the ancestral allelic form of HopZ1; therefore, this new evidence suggested that differentiated resistance systems have evolved in plant hosts to adapt to HopZ1 diversification in P. syringae.


First Report of Foliar Infection of Cabbage by Phytophthora drechsleri in Argentina.
Cabbage (Brassica oleracea var. capitata L.) is a popular crop grown along the northeast horticultural belt of Buenos Aires Province, Argentina. In the summer of 2010, fields in this region remained flooded for long periods due to frequent and intense precipitation (560 mm from January to March). Commercial cabbage crops in the cities of Luján and General Rodríguez developed patches of diseased plants that were stunted and wilted. Affected plants had necrotic areas in the crowns and roots. Symptoms expanded to the upper stems, leaving vascular tissues exposed. During April 2010, samples from 2 fields were brought to the laboratory where the stems were washed thoroughly and disinfected with a 1% bleach solution for 2 minutes. Small pieces (5 mm in diameter) were removed from the lesion edge, plated on V8 agar (V8A) plates, and incubated at 24°C in the dark for 5 days. Four isolates were transferred to V8A using hyphal tips. Morphological studies were performed on the V8A cultures as well as plates flooded with tap water. Sporangia were obpyriform, nonpapillate, persistent, and variable in size, averaging 44 × 28 μm. Each isolate belonged to the A1 mating type when paired with P. capsici tester isolates, CBS 370.72 and CBS 111.334 (Fungal Biodiversity Centre, CBS, Utrecht, the Netherlands). The isolates produced amphigynous antheridia, and chlamydospores were present but scarce. Maximum temperatures for growth (37°C) were also performed. Edited sequences of the internal transcribed spacer (ITS) region of the rDNA (GenBank Accession Nos. JQ653300, JQ653301, JQ653302, and JQ653303) were compared with Phytophthora sequences available in GenBank using the BLASTN search utility (1) and aligned to the data set of Cooke et al. (2). Sequences of the four isolates (strains 2: R-cai-cuello-col-3, 3: R-cai-cuello-col-18, 4: R-AN-col-1A and 5: R-AN-col-1B) matched 100% with GenBank sequences of P. drechsleri (100% coverage, 100% identity and no gaps). Based on these results, the four Argentinian cabbage isolates were identified as P. drechsleri (3). Pathogenicity tests were completed using three detached heads of mature cabbage plants (B. oleracea var. capitata) for each isolate. A 5-mm colonized mycelial plug of the appropriate isolate was placed on the main vein of the outermost leaves. For the control treatments, three heads were inoculated with non-colonized V8A plugs. The inoculated and control heads of cabbage were incubated in plastic boxes wrapped in black nylon bags at 24°C for 4 days. Broccoli (B. oleracea var. italica) and cauliflower (B. oleracea var. botrytis) were also tested following the same procedure. All heads of cabbage, broccoli, and cauliflower developed necrotic lesions 2 to 4 cm in diameter and a dark grey color. Control heads of each plant remained green. P. dreschleri was consistently reisolated as described above from the inoculated heads, but not from the control heads. To our knowledge, this is the first report of cabbage as a host for P. dreschleri in Argentina. Frezzi (4) reported this species as a pathogen of Chrysanthemum cinerariefolium, Celosia plumosa, Schinus molle, and Solanum lycopersicum in Argentina in 1950. References: (1) S. S. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. L. Cooke et al. Fungal Gen. Biol. 30:17, 2000. (3) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society Press, St. Paul, MN, 1996. (4) M. J. Frezzi. Rev. Invest. Agric. Buenos Aires 4:49, 1950.


Canadian dog owners' use and perceptions of cannabis products.
The legal market for recreational and medicinal cannabis for human consumption is growing worldwide. At the same time, marketing of cannabis products for use in pets is expanding. Yet, there is little research exploring the effects of cannabis use in veterinary medicine. This study used an anonymous, online survey to assess Canadian pet owners' reasons for purchasing cannabis products for their dogs, and their perceptions regarding efficacy of these treatments. Owners purchased cannabis products for treatment of pain, inflammation, and anxiety in dogs, and perceived these preparations to be equally or more effective than conventional medications. Most owners reported only minimal side effects in their dogs. Despite indicating comfort in discussing canine cannabis administration with their veterinarian, most owners relied on commercial websites for product information. The main reasons for choosing cannabis products were the ability to use as an adjuvant to other therapies, and the perception of it being a natural substance. Given this information, it is incumbent upon veterinarians to appropriately counsel their clients, and also to advocate for evidence-based studies to evaluate the efficacy of cannabis use in non-human species.


The long-term effects of Medicaid managed care on obstetrics care in three California counties.
To evaluate the long-term effects of Medicaid managed care (MMC) on obstetric service use and program costs in California. Longitudinal administrative data on Medi-Cal enrollment and claims and encounters related to pregnancy and delivery services were gathered from three counties--two long-standing MMC counties and one traditional fee-for-service Medicaid county--in California between 1987 and 1992. We studied Aid to Families with Dependent Children (AFDC) beneficiaries with live singleton vaginal deliveries with associated hospital stays of 14 days or less. Effects of managed care were examined with respect to prenatal visits, length of stay for delivery, maternal postpartum readmission rates, and total program expenditures. Multivariate analyses examined how the relative effect of managed care on service use and program expenditures in each MMC county evolves over time in comparison to fee-for-service. We controlled for length of Medi-Cal enrollment prior to delivery, data censoring, and individual characteristics such as race and age. Prenatal care use is consistently lower in the MMC counties, although all three counties' prenatal care provision is well below the national standard. Drastic increases in one-day-stay deliveries were found: up to almost 50 percent of deliveries in MMC counties were one-day stays. Program cost savings associated with MMC enrollment are unambiguous. MMC cost savings might have come at the expense of reduced provision of prenatal care and shorter delivery length of stay. Future studies should verify any possible causal link and the effects on maternal and infant health outcomes.


Deliberation in the motor system: reflex gains track evolving evidence leading to a decision.
Both decision making and sensorimotor control require real-time processing of noisy information streams. Historically these processes were thought to operate sequentially: cognitive processing leads to a decision, and the outcome is passed to the motor system to be converted into action. Recently, it has been suggested that the decision process may provide a continuous flow of information to the motor system, allowing it to prepare in a graded fashion for the probable outcome. Such continuous flow is supported by electrophysiology in nonhuman primates. Here we provide direct evidence for the continuous flow of an evolving decision variable to the motor system in humans. Subjects viewed a dynamic random dot display and were asked to indicate their decision about direction by moving a handle to one of two targets. We probed the state of the motor system by perturbing the arm at random times during decision formation. Reflex gains were modulated by the strength and duration of motion, reflecting the accumulated evidence in support of the evolving decision. The magnitude and variance of these gains tracked a decision variable that explained the subject's decision accuracy. The findings support a continuous process linking the evolving computations associated with decision making and sensorimotor control.


Haunted by a doppelgänger: irrelevant facial similarity affects rule-based judgments.
Judging other people is a common and important task. Every day professionals make decisions that affect the lives of other people when they diagnose medical conditions, grant parole, or hire new employees. To prevent discrimination, professional standards require that decision makers render accurate and unbiased judgments solely based on relevant information. Facial similarity to previously encountered persons can be a potential source of bias. Psychological research suggests that people only rely on similarity-based judgment strategies if the provided information does not allow them to make accurate rule-based judgments. Our study shows, however, that facial similarity to previously encountered persons influences judgment even in situations in which relevant information is available for making accurate rule-based judgments and where similarity is irrelevant for the task and relying on similarity is detrimental. In two experiments in an employment context we show that applicants who looked similar to high-performing former employees were judged as more suitable than applicants who looked similar to low-performing former employees. This similarity effect was found despite the fact that the participants used the relevant résumé information about the applicants by following a rule-based judgment strategy. These findings suggest that similarity-based and rule-based processes simultaneously underlie human judgment.


Family-based treatment for children and adolescents with eating disorders: a mixed-methods evaluation of a blended evidence-based implementation approach.
In this study, we evaluated a blended implementation approach with teams learning to provide family-based treatment (FBT) to adolescents with eating disorders. Four sites participated in a sequential mixed method pre-post study to evaluate the implementation of FBT in their clinical settings. The implementation approach included: (a) preparatory site visits; (b) the establishment of implementation teams; (c) a training workshop; (d) monthly clinical consultation; (e) monthly implementation consultation; and (f) fidelity assessment. Quantitative measures examining attitudes toward evidence-based practice, organizational learning environment and organizational readiness for change, as well as, individual readiness for change were delivered pre- and postimplementation. Correlational analyses were used to examine associations between baseline variables and therapist fidelity to FBT. Fundamental qualitative description guided the sampling and data collection for the qualitative interviews performed at the conclusion of the study. Seventeen individuals participated in this study (nine therapists, four medical practitioners, and four administrators). The predetermined threshold of implementation success of 80% fidelity in every FBT session was achieved by only one therapist. However, mean fidelity scores were similar to those reported in other studies. Participant attitudes, readiness, and self-efficacy were not associated with fidelity and did not change significantly from pre- to postimplementation. In qualitative interviews, all participants reported that the implementation intervention was helpful in adopting FBT. Our blended implementation approach was well received by participants. A larger trial is needed to determine which implementation factors predict FBT fidelity and impact patient outcomes.


Abdominal Aortic Calcifications Influences the Systemic and Renal Hemodynamic Response to Renal Denervation in the DENERHTN (Renal Denervation for Hypertension) Trial.
The DENERHTN (Renal Denervation for Hypertension) trial confirmed the efficacy of renal denervation (RDN) in lowering daytime ambulatory systolic blood pressure when added to standardized stepped-care antihypertensive treatment (SSAHT) for resistant hypertension at 6 months. This post hoc exploratory analysis assessed the impact of abdominal aortic calcifications (AAC) on the hemodynamic and renal response to RDN at 6 months. In total, 106 patients with resistant hypertension were randomly assigned to RDN plus SSAHT or to the same SSAHT alone (control group). Total AAC volume was measured, with semiautomatic software and blind to randomization, from the aortic hiatus to the iliac bifurcation using the prerandomization noncontrast abdominal computed tomography scans of 90 patients. Measurements were expressed as tertiles. The baseline-adjusted difference in the change in daytime ambulatory systolic blood pressure from baseline to 6 months between the RDN and control groups was -10.1 mm Hg (P=0.0462) in the lowest tertile and -2.5 mm Hg (P=0.4987) in the 2 highest tertiles of AAC volume. Estimated glomerular filtration rate remained stable at 6 months for the patients in the lowest tertile of AAC volume who underwent RDN (+2.5 mL/min per 1.73 m2) but decreased in the control group (-8.0 mL/min per 1.73 m2, P=0.0148). In the 2 highest tertiles of AAC volume, estimated glomerular filtration rate decreased similarly in the RDN and control groups (P=0.2640). RDN plus SSAHT resulted in a larger decrease in daytime ambulatory systolic blood pressure than SSAHT alone in patients with a lower AAC burden than in those with a higher AAC burden. This larger decrease in daytime ambulatory systolic blood pressure was not associated with a decrease in estimated glomerular filtration rate. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01570777.


Expression of TP53 mutation-associated microRNAs predicts clinical outcome in head and neck squamous cell carcinoma patients.
TP53 mutation is associated with decreased survival rate in head and neck squamous cell carcinoma (HNSCC) patients. We set out to identify microRNAs (miRNAs) whose expression associates with TP53 mutation and survival in HNSCC. We analyzed TP53 status by direct sequencing of exons 2 through 11 of a prospective series of 121 HNSCC samples and assessed its association with outcome in 109 followed-up patients. We carried out miRNA expression profiling on 121 HNSCC samples and 66 normal counterparts. miRNA associations with TP53 mutations and outcome were evaluated. A TP53 mutation was present in 58% of the tumors and TP53 mutations were significantly associated with a shorter recurrence-free survival. This association was stronger in the clinical subgroup of patients subjected to adjuvant therapy after surgery. The expression of 49 miRNAs was significantly associated with TP53 status. Among these 49, we identified a group of 12 miRNAs whose expression correlates with recurrence-free survival and a group of 4 miRNAs that correlates with cancer-specific survival. The two groups share three miRNAs. Importantly, miRNAs that correlate with survival are independent prognostic factors either when considered individually or as signatures. miRNAs expression associates with TP53 status and with reduced survival after surgical treatment of squamous cell carcinoma of the head and neck.


Impact of rotavirus vaccination on hospital-acquired rotavirus gastroenteritis in children.
Data show that after the implementation of routine rotavirus vaccination for infants in the United States, community-acquired (CA) rotavirus cases declined substantially in the 2007-2008 season. The impact of community-based rotavirus vaccination on the substantial burden of hospital-acquired (HA) rotavirus has not been documented. We assessed CA and HA rotavirus, respiratory syncytial virus, and influenza infections at Children's Memorial Hospital for 5 winter seasons (defined as occurring from September through May) from 2003 to 2008. We also report rotavirus data from the 2008-2009 season. A similar dramatic decline (>60% compared with the median of previous seasons) occurred in the rates of cases of both CA (P < .0001) rotavirus hospitalizations and HA (P < .01) rotavirus infections in the 2007-2008 season compared with previous seasons, whereas the rates of CA and HA influenza and respiratory syncytial virus, respectively, remained stable. Improvements in hand-hygiene compliance did not correlate with a reduction in the transmission rate of rotavirus in the hospital. Both CA and HA rotavirus rates remained much lower in the 2008-2009 than in the 2003-2007 seasons. Community-based rotavirus vaccination is associated with a substantial reduction in the number of children who are admitted with rotavirus. These data also indicate that routine community-based rotavirus infant vaccination protects hospitalized children from acquiring rotavirus. Vaccination efforts should be encouraged as a strategy to affect the substantial burden of HA rotavirus.


Validation of planning-free vessel-encoded pseudo-continuous arterial spin labeling MR imaging as territorial-ASL strategy by comparison to super-selective p-CASL MRI.
Vessel-encoded (VE) pseudo-continuous arterial spin labeling (p-CASL) is a territorial ASL (T-ASL) technique to identify the perfusion territories of cerebral arteries. The aim of this study was to validate the output of three Vessel-encoded p-CASL image processing methods, k-means clustering with and without subsequent linear analysis and a Bayesian framework, by comparison with the perfusion maps acquired with super-selective p-CASL. The comparison was done quantitatively using the Hausdorff distance and Dice similarity coefficient in the territories of the right and left internal carotid arteries, the basilar artery, and the right and left vertebral arteries. A qualitative comparison was done in the areas of the anterior and posterior circulation, and the deep gray matter. The overall agreement between the Vessel-encoded p-CASL image processing methods and super-selective p-CASL was good; with the difference that the linear analysis and the Bayesian framework were able to detect mixed perfusion. Planning-free Vessel-encoded p-CASL with k-means clustering appears suitable as a general purpose T-ASL strategy, but to determine mixed perfusion a combination with linear analysis, or the Bayesian framework is preferable, which are superior in this regard. To accurately determine the perfusion territory of a single vessel, super-selective p-CASL is still recommended.


Presynaptic localisation of the nicotinic acetylcholine receptor beta2 subunit immunoreactivity in rat nigrostriatal dopaminergic neurones.
Nicotinic acetylcholine receptors (nAChR) are widely distributed in the central nervous system, where they exert a modulatory influence on synaptic transmission. For the striatum, pharmacological evidence supports the presence of presynaptic alpha3beta2* and alpha4beta2* nAChR that modulate dopamine release from nigrostriatal terminals. The objective of this study was to examine the precise subcellular distribution of the nAChR beta2 subunit in these neurones and its localisation at presynaptic sites. Double immunolabelling with tyrosine hydroxylase (TH) at the confocal level revealed that the cell bodies and axon terminals (synaptosomes) of nigrostriatal neurones were also immunoreactive for the nAChR beta2 subunit. Double-preembedding electron microscopy confirmed that beta2-immunogold labelling was enriched in TH-positive terminals in the dorsal striatum. Quantitative analysis of doubly immunogold-labelled sections in postembedding electron microscopy showed that 86% of TH-positive axonal boutons are also labelled for the nAChR beta2 subunit, whereas 45% of beta2 subunit-immunolabeled boutons do not contain TH. Thus the beta2 subunit is localised within at least two populations of axon terminals in the dorsal striatum. In these structures, 15% of beta2 subunit immunoreactivity was at the plasma membrane but was rarely associated with synapses. These findings are compatible with functional presynaptic beta2-containing nAChR that may be stimulated physiologically by acetylcholine that diffuses from synaptic or nonsynaptic sites of acetylcholine release. These results demonstrate the presynaptic localisation of an nAChR subunit in nigrostriatal dopaminergic neurones, providing morphological evidence for the presynaptic nicotinic modulation of dopamine release.


LMO7 and LIMCH1 interact with LRIG proteins in lung cancer, with prognostic implications for early-stage disease.
The human leucine-rich repeats and immunoglobulin-like domains (LRIG) protein family comprises the integral membrane proteins LRIG1, LRIG2 and LRIG3. LRIG1 is frequently down-regulated in human cancer, and high levels of LRIG1 in tumor tissue are associated with favorable clinical outcomes in several tumor types including non-small cell lung cancer (NSCLC). Mechanistically, LRIG1 negatively regulates receptor tyrosine kinases and functions as a tumor suppressor. However, the details of the molecular mechanisms involved are poorly understood, and even less is known about the functions of LRIG2 and LRIG3. The aim of this study was to further elucidate the functions and molecular interactions of the LRIG proteins. A yeast two-hybrid screen was performed using a cytosolic LRIG3 peptide as bait. In transfected human cells, co-immunoprecipitation and co-localization experiments were performed. Proximity ligation assay was performed to investigate interactions between endogenously expressed proteins. Expression levels of LMO7 and LIMCH1 in normal and malignant lung tissue were investigated using qRT-PCR and through in silico analyses of public data sets. Finally, a clinical cohort comprising 355 surgically treated NSCLC cases was immunostained for LMO7. In the yeast two-hybrid screen, the two paralogous proteins LMO7 and LIMCH1 were identified as interaction partners to LRIG3. LMO7 and LIMCH1 co-localized and co-immunoprecipitated with both LRIG1 and LRIG3. Endogenously expressed LMO7 was in close proximity of both LRIG1 and LRIG3. LMO7 and LIMCH1 were highly expressed in normal lung tissue and down-regulated in malignant lung tissue. LMO7 immunoreactivity was shown to be a negative prognostic factor in LRIG1 positive tumors, predicting poor patient survival. These findings suggest that LMO7 and LIMCH1 physically interact with LRIG proteins and that expression of LMO7 is of clinical importance in NSCLC.


Caught in the middle: combined impacts of shark removal and coral loss on the fish communities of coral reefs.
Due to human activities, marine and terrestrial ecosystems face a future where disturbances are predicted to occur at a frequency and severity unprecedented in the recent past. Of particular concern is the ability of systems to recover where multiple stressors act simultaneously. We examine this issue in the context of a coral reef ecosystem where increases in stressors, such as fisheries, benthic degradation, cyclones and coral bleaching, are occurring at global scales. By utilizing long-term (decadal) monitoring programs, we examined the combined effects of chronic (removal of sharks) and pulse (cyclones, bleaching) disturbances on the trophic structure of coral reef fishes at two isolated atoll systems off the coast of northwest Australia. We provide evidence consistent with the hypothesis that the loss of sharks can have an impact that propagates down the food chain, potentially contributing to mesopredator release and altering the numbers of primary consumers. Simultaneously, we show how the effects of bottom-up processes of bleaching and cyclones appear to propagate up the food chain through herbivores, planktivores and corallivores, but do not affect carnivores. Because their presence may promote the abundance of herbivores, the removal of sharks by fishing has implications for both natural and anthropogenic disturbances involving the loss of corals, as herbivores are critical to the progress and outcome of coral recovery.


A systematic review of techniques and effects of self-help interventions for tinnitus: Application of taxonomies from health psychology.
Self-help interventions are followed by people independently with minimal or no therapist contact. This review aims to assess the effectiveness of self-help interventions for adults with chronic tinnitus and systematically identify the self-help techniques used. Systematic review and application of health psychology taxonomies. Electronic database searches were conducted, supplemented by citation searching and hand-searching of key journals. Prospective controlled trials, which used measures of tinnitus distress, functional management, anxiety, depression, and quality of life, were included. Michie et al's behaviour change techniques (BCTs) taxonomy and Taylor et al's PRISMS taxonomy of self-management components were applied to describe interventions. Five studies were included, providing low-to-moderate levels of evidence. Randomized controlled trial studies were too few and heterogeneous for meta-analysis to be performed. Studies comparing self-help interventions to therapist-guided interventions and assessing non tinnitus-specific psychosocial outcomes and functional management were lacking. Fifteen BCTs and eight self-management components were identified across interventions. A lack of high-quality and homogeneous studies meant that confident conclusions could not be drawn regarding the efficacy of self-help interventions for tinnitus. Better reporting and categorization of intervention techniques is needed for replication in research and practice and to facilitate understanding of intervention mechanisms.


Genome-wide trans-ancestry meta-analysis provides insight into the genetic architecture of type 2 diabetes susceptibility.
To further understanding of the genetic basis of type 2 diabetes (T2D) susceptibility, we aggregated published meta-analyses of genome-wide association studies (GWAS), including 26,488 cases and 83,964 controls of European, east Asian, south Asian and Mexican and Mexican American ancestry. We observed a significant excess in the directional consistency of T2D risk alleles across ancestry groups, even at SNPs demonstrating only weak evidence of association. By following up the strongest signals of association from the trans-ethnic meta-analysis in an additional 21,491 cases and 55,647 controls of European ancestry, we identified seven new T2D susceptibility loci. Furthermore, we observed considerable improvements in the fine-mapping resolution of common variant association signals at several T2D susceptibility loci. These observations highlight the benefits of trans-ethnic GWAS for the discovery and characterization of complex trait loci and emphasize an exciting opportunity to extend insight into the genetic architecture and pathogenesis of human diseases across populations of diverse ancestry.


Interleukin 10 (IL-10) and viral IL-10 strongly reduce antigen-specific human T cell proliferation by diminishing the antigen-presenting capacity of monocytes via downregulation of class II major histocompatibility complex expression.
Interleukin 10 (IL-10) and viral IL-10 (v-IL-10) strongly reduced antigen-specific proliferation of human T cells and CD4+ T cell clones when monocytes were used as antigen-presenting cells. In contrast, IL-10 and v-IL-10 did not affect the proliferative responses to antigens presented by autologous Epstein-Barr virus-lymphoblastoid cell line (EBV-LCL). Inhibition of antigen-specific T cell responses was associated with downregulation of constitutive, as well as interferon gamma- or IL-4-induced, class II MHC expression on monocytes by IL-10 and v-IL-10, resulting in the reduction in antigen-presenting capacity of these cells. In contrast, IL-10 and v-IL-10 had no effect on class II major histocompatibility complex (MHC) expression on EBV-LCL. The reduced antigen-presenting capacity of monocytes correlated with a decreased capacity to mobilize intracellular Ca2+ in the responder T cell clones. The diminished antigen-presenting capacities of monocytes were not due to inhibitory effects of IL-10 and v-IL-10 on antigen processing, since the proliferative T cell responses to antigenic peptides, which did not require processing, were equally well inhibited. Furthermore, the inhibitory effects of IL-10 and v-IL-10 on antigen-specific proliferative T cell responses could not be neutralized by exogenous IL-2 or IL-4. Although IL-10 and v-IL-10 suppressed IL-1 alpha, IL-1 beta, tumor necrosis factor alpha (TNF-alpha), and IL-6 production by monocytes, it was excluded that these cytokines played a role in antigen-specific T cell proliferation, since normal antigen-specific responses were observed in the presence of neutralizing anti-IL-1, -IL-6, and -TNF-alpha mAbs. Furthermore, addition of saturating concentrations of IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha to the cultures had no effect on the reduced proliferative T cell responses in the presence of IL-10, or v-IL-10. Collectively, our data indicate that IL-10 and v-IL-10 can completely prevent antigen-specific T cell proliferation by inhibition of the antigen-presenting capacity of monocytes through downregulation of class II MHC antigens on monocytes.